PHARCHEM2 LAB

CYANOGENIC GLYCOSIDES
3CPH Himelo, Alexis Hui, Hans Gerard Jariel, Jesse Lance Lim, Hannah Lorraine Magana, Johnvee Mamburam, Liezel

DEFINITION
Also known as cyanogenetic, cyanophore, or

cyanogenic glycosides N-containing secondary metabolites whose biosynthetic origin is similar to that of glucosinolates O-glycosides yielding HCN gas on hydrolysis

DEFINITION
Intermediately polar, water-soluble compounds that

often accumulate in the vacuoles of plant cells (O-glycosides of cyanohydrins: -hydroxynitriles) Formed in the cytoplasm Storage at the epidermal vacuoles in sorghum -glucosidase degrade cyanogenic glycoside

DEFINITION
A level of 10mg HCN per kg is the minimum for a plant

to be considered cyanogenic Minimal lethal dose of HCN 0.5-3.5mg per kg body weight

DEFINITION
Important families in which cyanogenic glycoside occurs in :

Araceae Asteraceae Euphorbiacee Fabaceae Flacourtiaceae Maleessherba

Proteaceae Rosaceae Sapindaceae Turneraceae Compositae Gramineae Leguminoseae

DEFINITION
cyanoglycosides account for approximately 90% of the

wider group of plant toxins known as cyanogens

The key characteristic of these toxins is cyanohydrins

that have been established by glycosylation, or the attachment of sugar, to form cyanogenic glycosides.

DEFINITION
Cyanogenesis- production of hydrocyanic acid (HCN) Presence of cyanide makes the plant toxic to animals

Plant tissue is damaged (herbivory, trampling, intense

heat, or frost ), cyanide is released

DEFINITION
Hydrolysis is accomplished by -glucosidase,

producing sugars, cyanohydrin (spontaneously decomposes to HCN and ketone or aldehyde) Explains the defense mechanism of the plant against predators plant tissue is damaged, cyanogenic glycoside and enzymes (in the cytoplasm) are put in contact and cyanohydric acid is released.

DEFINITION
remove the sugar part of the molecule and release

toxic hydrogen cyanide. Storing them in inactive forms in the cytoplasm prevents them from damaging the plant under normal conditions.

GENERAL STRUCTURE

Aglycone: -hydronitrile derivatives Sugar moeity: D-glucose (exceptions amygdalin, vicianin,

lucumin, linustatin, neolinustatin - have disaccharides) R1 and R2 : aliphatic or aromatic substituent (or hydrogen) Carbinolcarbon of the aglycone is usually chiral since R1 and R2 are not identical

DEFINITION
All known cyanogenic glycosides are -linked, mostly with

D-glucose.
Cyanogenic glycosides play pivotal roles in organization of

chemical defense system in plants and in plantinsect interactions.

MEDICINAL/ PHARMACOLOGICAL USE

Sedative (HCN content) Vasodilator Chemotherapy for cancer Tuberculosis Leprosy Diarrhea Lowers cholesterol Demulcent Respiratory inhibitor

Increase bleeding time Constipation Gastritis Anti- inflammatory Vitamin B17 Antitussive Lowers blood pressure Anti- spasmodic

TRADITIONAL USE
Cassava plant and Bamboo Shoots

Linamarin and taxiphyllin

-

Traditionally used as food treating snakebites, boils, inflammation, cancer (cassava plant) Traditionally used to make older antineoplastic drugs

Amygdalin, Linamarin
-

CLASSIFICATION BASED ON BIOSYNTHETIC ORIGIN

1. Phenylalanine 2. Tyrosine 3. Leucine 4. Valine and Isoleucine 5. Cyclopentenylglyci ne 7.Nicotinic Acid 8.Nitroacids and nitroalcohol 9.Tyrosine Nitrile glycoside

DERIVED FROM PHENYLALANINE

Derived from L-phenylalanine Found in Rosidae and Asteridae

Amygdalin (R)-Prunasin (S)-Sambunigrin (R)-Vicianin (R)-Lucumin

PHENYLALANINE: AMYGDALIN

IUPAC NAME:
2-phenyl-2-[(2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-[[(2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6(hydroxymethyl)tetrahydropyran-2-yl]oxymethyl]tetrahydropyran- 2-yl]oxy-acetonitrile

Wide-spread in seeds of the Rosaceae (apples and peaches) Use: Flavoring agent, anti-cancer properties

PHENYLALANINE: AMYGDALIN
Colorless crystalline bitter glycoside Anticancer claim: Control of sickle cell

anemia not recognized by FDA as a cancer drug because of high HCN content, potential for overdose or poisoning pure amygdalin will break down when taken internally into glucose, benzaldehyde and HCN (poisonous)

PHENYLALANINE: AMYGDALIN
Beta-glucosidase is the enzyme that initiates the

release of cyanide from amygdalin. It is found in the small intestine. This can lead to an unpredictable and potentially lethal toxicity when pure amygdalin is taken orally Hydrocyanic acid: VERY POISONOUS AND MAKES THE DRUG UNSUITABLE FOR INTERNAL CONSUMPTION

PHENYLALANINE: (R)-PRUNASIN
IUPAC name:
(2R)-2-phenyl-2-[(2R,3R,4S,5S,6R)-3,4,5trihydroxy-6-(hydroxymethyl) tetrahydropyran-2-yl]oxy-acetonitrile

found in various plants of the

genus Prunus Expectorant, sedative, antispasmodic effect and digestive properties (small quantities) Obtained by partial hydrolysis of amygdalin by enzyme prunase

PHENYLALANINE: (S)-SAMBUNIGRIN

IUPAC name:
(2S)-2-phenyl-2-[(2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6(hydroxymethyl)tetrahydropyran-2-yl]oxy-acetonitrile

In barks, leaves, unripe berries, seeds of ripe berries

PHENYLALANINE: (R)-VICIANIN

IUPAC name:
(2R)-2-phenyl-2-[(2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-[[(2S,3R,4S,5S)3,4,5-trihydroxyoxan-2-yl]oxymethyl]oxan-2-yl]oxyacetonitrile

In seed of the common vetch (Vicia sativa)

PHENYLALANINE: (R)-LUCUMIN

IUPAC name:
2-phenyl-2-[(2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-[[(2S,3R,4S,5R)-3,4,5trihydroxyoxan-2-yl]oxymethyl]oxan-2-yl]oxyacetonitrile

Occurs in the seed of Colocarpum sapota (Sapotaceae) Sugar moiety: primverose

DERIVED FROM TYROSINE

Commonly occurs in monocotyledonous angiosperms

and Magnolidae

Dhurrin Triglochinin

Proteacin

TYROSINE: (S)-DHURRIN

IUPAC name
(2S)-2-(4-Hydroxyphenyl)-2-[[(2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)-2tetrahydropyranyl]oxy]acetonitrile

Make up to 30% of the dry weight of the leaves and coleoptiles of etiolated sorghum seedlings (Poaceae)

TYROSINE: TRIGLOCHININ

IUPAC name:
4-[cyano-[3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxymethylidene]hex-2-enedioic acid

Found in edible aroids of Taro leaves and stem

TYROSINE: (S)- PROTEACIN

IUPAC name:
(2S)-2-[(2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl] oxy-2-[4[(2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyphenyl]acetonitrile

Only known from the seed kernels of bitter Macadamia nuts, Macadamia ternifolia (Proteaceae)

DERIVED FROM LEUCINE

HAVE RESTRICTED DISTRIBUTION (R)-Epiproacacipetalin (S)-Proacaciberin (S)-Heterodendrin (R)-Epiheterodendrin

LEUCINE: (R)-EPIPROACACIPETALIN

IUPAC name:
(2R)-3-methyl-2-[(2R,3R,4S,5S,6R)-3,4, 5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxybut-3-enenitrile

Found in Acacia sp. with Proacaciberin

LEUCINE: (S)-PROACACIBERIN

IUPAC name:
(2S)-3-methyl-2-[(2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-[[(2S,3R,4S,5S)-3, 4,5-trihydroxyoxan-2-yl]oxymethyl]oxan-2-yl]oxybut-3-enenitrile

Sugar moiety: vicianose unit

LEUCINE: (S)-HETERODENDRIN

IUPAC name:
(2S)-3-methyl-2-[(2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl) oxan-2-yl]oxybutanenitrile

only found in Sapindaceae and Poaceae

LEUCINE: (R)-EPIHETERODENDRIN

IUPAC name:
(2R)-3-methyl-2-[(2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl) oxan-2-yl]oxybutanenitrile

only found in Sapindaceae and Poaceae

DERIVED FROM VALINE & ISOLEUCINE

Linamarin (R)-Lotaustarin Linustatin Neolinustatin Sarmentosin Epoxide

VALINE & ISOLEUCINE: LINAMARIN

IUPAC name: 2-methyl-2-[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl] oxy-propanenitrile Linum usitatissimum (Linaceae) Use: treatment of diarrhea, constipation, gastritis; poultice skin

inflammation

VALINE & ISOLEUCINE: (R)LOTAUSTRALIN

IUPAC name:
(2R)-2-methyl-2-{[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro2H-pyran-2-yl]oxy}butanenitrile

almost always co-occur with Linamarin Structurally related to Linamarin Glucoside of methyl ethyl ketone cyanohydrin

VALINE & ISOLEUCINE: LINUSTATIN

IUPAC name:
2-(6-O--D-Glucopyranosyl--D-glucopyranosyloxy)-2-methylpropanenitrile

gentibiose derivative; found in flaxseed and Rossiflora sp.

VALINE & ISOLEUCINE: NEOLINUSTATIN

IUPAC name:
(2R)-2-(6-O--D-Glucopyranosyl--D-glucopyranosyloxy)-2-methylbutanenitrile

gentibiose derivative; found in flaxseed and Rossiflora sp.

VALINE & ISOLEUCINE: SARMENTOSIN EPOXIDE

IUPAC name:
2-(hydroxymethyl)-3-[[(2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6(hydroxymethyl)oxan-2-yl]oxymethyl]oxirane-2-carbonitrile

species of Asteraceae, Euphorbiaciae, Fabaceae and Linaceae Contains an epoxide ring and is not a glycoside of alpha-nitrile Opening of epoxide ring results to liberation of cyanide, spontaneously

DERIVED FROM CYCLOPENTENYLGLYCINE

Contains a cyclopentanoid ring structure Derived from non-protein amino acid 2-(2-

cyclopentenyl)glycine

(1R)-Deidaclin (1R,4R)-Volkenin Gynocardin

CYCLOPENTENYLGLYCINE: (1R)- DEIDACLIN

IUPAC name:
(1R)-1-[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6- (hydroxymethyl)oxan-2yl]oxycyclopent-2-ene-1-carbonitrile

leaves of Deidamia clematoides

CYCLOPENTENYLGLYCINE: (4R)-VOLKENIN

IUPAC name:
(4R)-4-hydroxy-1-[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl) oxan-2-yl]oxycyclopent-2-ene-1-carbonitrile

Also known as epitetraphyllin B

CYCLOPENTENYLGLYCINE: GYNOCARDIN

IUPAC name:
4,5-dihydroxy-1-[3,4,5-trihydroxy-6(hydroxymethyl)oxan-2-yl] oxycyclopent-2-ene-1-carbonitrile

Gynocardia sp. and Pangium sp.

NICOTINIC ACID: ACALYPHIN

IUPAC name:
2-hydroxy-4-methoxy-1-methyl-6-oxo-3-[3,4,5-trihydroxy-6(hydroxymethyl)oxan-2-yl]oxy-2H-pyridine-3-carbonitrile

from Acalypha indica from Euphorbiaceae

DERIVED FROM NITROGLYCOSIDES

nitroacids and nitroalcohol 3-nitropropionic acid is derived from

aspartic acid few poisonous compounds to several insect and other animals are derived from 3-nitro-1-propanol

DERIVED FROM NITRILE GLYCOSIDES

Noncyanogenic compounds Structural similarities to intermediates in cyanogenic

glycoside biosynthesis

Simmondsin

NITRILE GLYCOSIDE: SIMMONDSIN

IUPAC name: 2-(cyanomethylene)-3-hydroxy-4,5- dimethoxycyclohexyl-beta-D-glucoside from the jojoba plant (Simmondsia chinensis) Appetite suppressant effect (obese individuals) Traditionally though to be toxic substance due to jojoba seed causing weight loss

EXTRACTION PROCEDURE I: MACERATION

1.Prepare 1 gram of ground leaves. Place it in a 1-liter beaker. 2.Prepare 1 liter of 95% ethanol. Carefully pour it in the beaker

with the ground leaves. Make sure it is properly soaked. 3.Stand for 48 hours. 4.After 48 hours, filter and collect the filtrate. 5.The filtrate is now ready for distillation.
*maceration allows the enzyme beta-glucosidase and the cyanogenic glycoside put in contact and cyanohydric acid is released

EXTRACTION PROCEDURE II: DISTILLATION

1. Assemble the distillation set-up. 2. Fill the distilling flask with the

extract. 3. The thermometer is inserted in the distilling flask through a hole in the cork stopper. The arm of the flask is inserted through a hole in the stopper of the condenser. 4. Heat the distilling flask using water bath.

EXTRACTION PROCEDURE II: DISTILLATION

5. Maintain the temperature with

the boiling point of ethanol, which is 80oC. 6. When almost all of the ethanol has already vaporized and the extract is semi-solid, remove the distilling flask from the heat source before all of the liquid is vaporized. 7. Collect the concentrated extract on the distilling flask.

EXTRACTION PROCEDURE: SOXHLET EXTRACTION

Requires large amount of

hazardous organic solvent ADVANTAGE: large amounts of drugs can be extracted with a much smaller quantity of solvent DISADVANTAGE: time consuming Makes the procedure excessive

EXTRACTION PROCEDURE: SOXHLET EXTRACTION

1. The grind sample prepared by the lab

assistant is weighed on an analytical balance. 2. The sample is mixed wit anhydrous sodium sulfate using a ratio of 4:1. 3. 150mL of hexane is poured into 250mL round- bottomed flask with boiling chips. 4. Soxhlet apparatus is set for the water batch of the concentrator apparatus.

EXTRACTION PROCEDURE: SOXHLET EXTRACTION

5. After an hour of extraction, the

round-bottomed flask is heated in the water batch of the concentrator apparatus. 6. The solvent is removed and excess water outside the flask is dried. 7. The extracted fat is weight and the fat content of the sample is calculated

PLANT SAMPLE: WILD CHERRY

Stem bark of Prunus serotina Prunasin

(cough remedies) sedative expectorant Antitussive Astringent Bronchitis Whooping cough Antispasmodic or Tonic effect Amygdalin Anti-cancer properties

PLANT SAMPLE: BITTER ALMOND

Seed of Prunus amygdalus Amygdalin

Sedative

Demulcent
INDUSTRIAL USES:

Used in perfumery and in the form of

liquors

PLANT SAMPLE: LINSEED

Dried fully riped seed of Linum usitatissimum Linamarin

Sedative effect on the respiratory system Anti-inflammatory Demulcent

PLANT SAMPLES: APRICOT PITS

Kernel of various varieties of Prunus americana

(Rosaceae) sold as a health food as source of laetrile or amygdalin

sold due to amygdalin but banned by

FDA due to ineffectivity and potential danger

PLANT SAMPLES

PLANT SAMPLES

WAYS OF DETOXIFYING PLANT FROM CYANIDE

Peeling Chopping Grating Soaking Boiling or Cooking Ensiling Drying

HUMAN HEALTH EFFECTS

Clinical symptoms of acute cyanide poisoning:

rapid respiration, drop in blood pressure, rapid pulse, headache, dizziness, vomiting, diarrhea, mental confusion, stupor, blue discoloration of the skin due to lack of oxygen (cyanosis), twitching and convulsions

HUMAN HEALTH EFFECTS

Cases of acute poisoning have been associated with

misuse, particularly of preparations from apricot pits, bitter almonds and cyanide rich apple seeds

HUMAN HEALTH EFFECTS

Cyanide is detoxified in the body, by the enzyme

rhodanase in the presence of sulphur- containing amino acids, to produce thiocyanate Goiter and cretinism due to iodine deficiency can be exacerbated by chronic consumption of insufficiently processed cassava

NEUROLOGICAL EFECTS
Konzo or spastic paraparesis is a motor neuron

disease characterized by irreversible weakness in the legs (speech and arms may be affected in severe cases) associated with a high and sustained intake of cassava (Manihot esculenta) in combinaton with a low protein intake

BIBLIOGRAPHY

BOOKS Barnes, et al. (2004). Fundamentals of Pharmacognosy and Phytotherapy. Important Natural Products and Phytomedicines Used Pharmacy and Medicine, Elsevier Science Limited. INTERNET Cyanogenic Glycosides (n.d.). Retrieved July 21, 2013from http://www.foodsafety.govt.nz/elibrary/industry/Cyanogenic_Gl ycosidesToxin_Which.pdf Vetter, J (2000). Plant Cyanogenic Glycosides. 38(1), 11-36. Retrieved July 2013 from http://www.ncbi.nlm.nih.gov/pubmed/10669009 22,

in

Speijers, G. (n.d.). Cyanogenic Glycosides. Retrieved July 23, 2013 from http://www.inchem.org/documents/jecfa/jecmono/v30je18.htm Khalid, M.A. (2011). Cyanophore (Cyanogenic) Glycosides. Retrieved July 2013 from Scribd.com/mobile/doc/76919142

22,

BIBLIOGRAPHY
Glycosides (n.d.). Retrieved July 22, 2013from http://quizlet.com/7619327/ubdr-zphytopharm-glycosides-flash-cards/ Adnan, A.A. (n.d.). LAB 1:FAT & OIL FROM OIL-SEEDS: PART I SOXHLET
EXTRACTION. Retrieved July 21, 2013 from

http://www.academia.edu/2967956/FAT_and_OIL_FROM_OILSEEDS_PART_I_SOXHLET_EXTRACTION Palmer M.E., Betz J.M. (2011). Chapter 118. Plants. In M.E. Palmer, J.M. Betz (Eds),Goldfrank's Toxicologic Emergencies, 9e. Retrieved July 24, 2013 http://www.accesspharmacy.com/content.aspx?aID=6529928. Cyanogenic Glycosides in Cassava and Bamboo Shoots (2004). Retrieved July 22, 2013 from http://www.foodstandards.gov.au/publications/documents/28_Cyan ogenic_glycosides.pdf